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KMID : 0603820090150040265
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Journal of Experimental & Biomedical Science
2009 Volume.15 No. 4 p.265 ~ p.272
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Optimization of Gene Delivery Mediated by Lipoplexes and Electroporation into Mouse Mesenchymal Stem Cells
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Kim Jong-Chul
Kim Hong-Sung
Lee Yeon-Kyung
Kim Jung-Seok
Park Sang-Il
Jung Hwa-Yeon
Park Yong-Serk
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Abstract
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Recently, mesenchymal stem cells (MSCs) began to be utilized as a vehicle for ex vivo gene therapy based on their plasticity. Effective and safe transfection of therapeutic genes is a critical step for genetic modification of MSCs. Therefore, optimization of in vitro gene delivery into MSCs is essential to provide genetically modified stem cells. In this study, various cationic liposomes, O,O’-dimyristyl-N-lysyl aspartate (DMKD), DMKD/cholesterol, O,O’-dimyristyl- N-lysyl glutamate (DMKE), DMKE/cholesterol, and N-[1-(2,3-dioleoyloxy)]-N,N,N-trimethylammonium propane methyl sulfate (DOTAP)/cholesterol, were mixed with plasmid DNA encoding luciferase (pAAV-CMV-Luc) at varied ratios, and then used for transfection to MSCs under varied conditions. The MSCs were also transfected by electroporation under varied conditions, such as voltage, pulse length, and pulse interval. According to the experimental results, electroporation-mediated transfection was more efficient than cationic liposome-mediated transfection. The best MSC transfection was induced by electroporation 3 times pulses for 2 ms at 200 V with 10 seconds of a pulse interval.
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KEYWORD
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Mouse mesenchymal stem cells, Cationic liposome, Electroporation, Gene delivery
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